Advanced Healthcare Materials
·
2023
·
IF 10
Ni Ding,Gangjian Luo,Huiting Li,Chengyuan Xing,Yuanji Gao,Wenjie Xi,Weijie Wu,Dan Wang,Lei Zheng,Yang Kang,Xinjin Chi
The mortality rate of sepsis remains high despite improvements in the diagnosis and treatment of sepsis using symptomatic and supportive therapies, such as anti-infection therapy and fluid resuscitation. Nucleic acid-based drugs have therapeutic potential, although their poor stability and low delivery efficiency have hindered their widespread use. Herein, it is confirmed that miR-223 can polarize proinflammation M1 macrophages to anti-inflammation M2 macrophages. A pH-sensitive nano-drug delivery system comprising β-cyclodextrin-poly(2-(diisopropylamino)ethyl methacrylate)/distearoyl phosphoethanolamine-polyethylene glycol (β-CD-PDPA/DSPE-PEG) is synthesized and developed to target M1 macrophages and miR-223 is encapsulated into nanoparticles (NPs) for sepsis treatment. NPs/miR-223 demonstrated in vitro pH responsiveness with favorable biosafety, stability, and high delivery efficiency. In vivo studies demonstrate that NPs/miR-223 are preferentially accumulated and retained in the inflammation site, thereby reducing inflammation and improving the survival rate of mice with sepsis while exhibiting ideal biosafety. Mechanically, NPs/miR-223 regulates macrophage polarization by targeting Pknox1 and inhibiting the activation of the NF-κB signaling pathway, thereby achieving an anti-inflammatory effect. Collectively, it is demonstrated that the miRNA delivery vector described here provides a new approach for sepsis treatment and accelerates the advancement of nucleic acid drug therapy.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MOLECULAR HUMAN REPRODUCTION
·
2018
·Vol. 25
·期 1
·5-16
·
IF 3.449
Sun Huihui,Li Dong,Yuan Ming,Li Qiuju,Zhen Qianwei,Li Ni,Wang Guoyun
STUDY QUESTIONDo exosomes play a role in the pathogenesis of endometriosis in a murine model?SUMMARY ANSWERExosomes from endometriosis (EMS) can alternatively activate macrophages and thus contribute to the development of lesions in mice.WHAT IS KNOWN ALREADYThe pathogenesis of endometriosis, an inflammatory disease, possibly involves peritoneal macrophages. Exosomes are recognized as a new communicator among cells and a key modulator in several inflammatory diseases.STUDY DESIGN, SIZE, DURATIONWe performed in vitr and in vivo experiments to demonstrate the role of exosomes in modulating macrophages. RAW264.7 cells (macrophages) were used to examine the effects of exosomes on macrophages in vitro. An experiment was also conducted in vivo, as follows. Fifty C57BL/6 female mice were randomly allocated to five control and five experimental groups (n = 5/group). The experimental group was injected i.p. with EMS-exosomes derived from eutopic stromal cells, starting on Day-7 then every day for 1 week. The control group received CON-exosomes from mice without endometriosis. Peritoneal macrophages were assessed over the next 6 days. On Day 0, all mice were injected i.p. with endometrium to establish the endometriosis model. On Day 14, all mice were sacrificed, ectopic lesions were counted and measured.PARTICIPANTS/MATERIALS, SETTING, METHODSExosomes were isolated from endometrial stromal cells (ESCs) by ultracentrifugation and characterized through transmission electron microscopy, nanoparticle tracking analysis and western blot. After treatment with exosomes, the polarization and phagocytic ability of the macrophages were detected by flow cytometry analysis, immunofluorescent staining and RT-PCR. C57BL/6 mice were utilized to establish an endometriosis model by i.p. injection of endometrial segments.MAIN RESULTS AND THE ROLE OF CHANCEAfter treatment with EMS-exosomes, the macrophages were polarized into an M2-like phenotype and their phagocytic ability decreased (P < 0.05 versus treatment with CON-exosomes). The total weight and volume of the lesions in mice treated with EMS-exosomes significantly increased compared with those in mice treated with CON-exosomes (P < 0.05). The infiltration of M2-like macrophages was enhanced in the EMS-exosome group (P < 0.001 versus treatment with CON-exosomes).LARGE SCALE DATAN/ALIMITATIONS, REASONS FOR CAUTIONDetection of endometriosis following exosome treatment was only performed in a murine endometriosis model. Clinical data and additional mechanism studies must be conducted to understand the role of exosomes in the pathogenesis of endometriosis.WIDER IMPLICATIONS OF THE FINDINGSThis study emphasizes the importance of EMS-exosomes in the pathogenesis of endometriosis. Further investigations on the exosome signaling pathways may contribute to the development of effective treatments for endometriosis.STUDY FUNDING/COMPETING INTEREST(S)This research was supported by grants (Nos. 81571417 and 81771552) from the National Science Foundation of China. The authors report no conflict of interest.
研究领域:
生殖生物学
·
免疫学
·
妇科学
✓ 已验证
DOI
PubMed
高影响因子
Cancer Cell International
·
2021
·Vol. 21
·期 1
·1-13
·
IF 5.722
Lei Jie,Chen Peng,Zhang Feng,Zhang Na,Zhu Jianfei,Wang Xiaoping,Jiang Tao
Background Exosomes are known to transmit microRNAs (miRNAs) to affect cancer progression, while the role of M2 macrophages-derived exosomes (M2 exosomes) conveying miR-501-3p in lung cancer (LC) remains unknown. We aim to explore the role of exosomal miR-501-3p in LC development via targeting WD repeat domain 82 (WDR82). Methods Lung cancer tissue and normal tissue specimens were collected, in which tumor-associated macrophages (TAM) were measured by immunohistochemistry. M2 macrophages were induced and treated with altered miR-501-3p, and then the exosomes were extracted and identified. MiR-501-3p and WDR82 expression in LC tissues and cell liens was determined. The predictive role of miR-501-3p in prognosis of LC patients was assessed, and the proliferation, colony formation ability, invasion, migration and apoptosis of the LC cells were determined. Targeting relationship between miR-501-3p and WDR82 was confirmed. Results TAM level was elevated in lung cancer tissues. MiR-501-3p was upregulated while WDR82 was downregulated in LC tissues and cell lines, and the M2 exosomes further upregulated miR-501-3p. M2 exosomes and exosomal miR-501-3p promoted LC cell growth. MiR-501-3p inhibition reversed the effect of M2 exosomes on LC cells. WDR82 was confirmed as a target gene of miR-501-3p. Conclusion M2 macrophages-derived exosomal miR-501-3p promotes the progression of LC via downregulating WDR82.
研究领域:
肿瘤学
·
分子生物学
·
细胞生物学
✓ 已验证
DOI
PubMed
高影响因子
Acta Biomaterialia
·
2024
·
IF 9.4
Shue Jin,Jing Wen,Yao Zhang,Ping Mou,Zeyu Luo,Yongrui Cai,Anjin Chen,Xiaoxue Fu,Weikun Meng,Zongke Zhou,Jidong Li,Weinan Zeng
Designing scaffolds that can regulate the innate immune response and promote vascularized bone regeneration holds promise for bone tissue engineering. Herein, electrospun scaffolds that combined physical and biological cues were fabricated by anchoring reparative M2 macrophage–derived exosomes onto topological pore structured nanofibrous scaffolds. The topological pore structure of the fiber and the immobilization of exosomes increased the nanoscale roughness and hydrophilicity of the fibrous scaffold. In vitro cell experiments showed that exosomes could be internalized by target cells to promote cell migration, tube formation, osteogenic differentiation, and anti-inflammatory macrophage polarization. The activation of fibrosis, angiogenesis, and macrophage was elucidated during the exosome-functionalized fibrous scaffold–mediated foreign body response (FBR) in subcutaneous implantation in mice. The exosome-functionalized nanofibrous scaffolds also enhanced vascularized bone formation in a critical-sized rat cranial bone defect model. Importantly, histological analysis revealed that the biofunctional scaffolds regulated the coupling effect of angiogenesis, osteoclastogenesis, and osteogenesis by stimulating type H vessel formation. This study elaborated on the complex processes within the cell microenvironment niche during fibrous scaffold–mediated FBR and vascularized bone regeneration to guide the design of implants or devices used in orthopedics and maxillofacial surgery. Statement of significance How to design scaffold materials that can regulate the local immune niche and truly achieve functional vascularized bone regeneration still remain an open question. Here, combining physical and biological cues, we proposed new insight to cell-free and growth factor-free therapy, anchoring reparative M2 macrophage–derived exosomes onto topological pore structured nanofibrous scaffolds. The exosomes functionalized-scaffold system mitigated foreign body response, including excessive fibrosis, tumor-like vascularization, and macrophage activation. Importantly, the biofunctional scaffolds regulated the coupling effect of angiogenesis, osteoclastogenesis, and osteogenesis by stimulating type H vessel formation.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
RSC Advances
·
2026
·
IF 6.1
Min Zou,Yunlong Liu,Ting Luo,Hui Yang,Jie Liang,Lang Tian,Jiachen Sun,Mufan Li
Background : severe bone defects caused by trauma or tumor resection markedly reduce quality of life. Although autologous bone grafting remains the clinical gold standard, donor-site morbidity restricts its broader use. Silk fibroin/nano-hydroxyapatite (SF/nHA) composites are promising for bone tissue engineering, but poor mechanical strength and limited regulation of the immune microenvironment remain major barriers to application. Methods : a graphene oxide (GO)-modified SF/nHA scaffold was developed as a dual-delivery system for interleukin-4 (IL-4) and vascular endothelial growth factor (VEGF). In vitro , macrophages and endothelial progenitor cells (EPCs) were cultured separately with the scaffolds to assess immunomodulatory and angiogenic effects. In vivo , scaffold degradation and bone repair were examined in rat subcutaneous implantation and calvarial defect models. Results : the GO-SF/nHA scaffold showed improved compressive modulus and stable biphasic release behavior. In vitro , sustained IL-4 promoted macrophage polarization toward the M2 phenotype, whereas VEGF enhanced EPC angiogenic activity. In vivo , dual delivery was associated with faster scaffold degradation and increased early neovascularization after subcutaneous implantation. In the critical-sized calvarial defect model, the I + V scaffold promoted macrophage M2 polarization and angiogenesis in parallel. At 8 weeks, this response was accompanied by greater collagen deposition and bone formation, with nearly complete defect bridging. Conclusion : co-delivery of IL-4 and VEGF enabled the GO-modified SF/nHA scaffold to promote macrophage M2 polarization, angiogenesis, and subsequent bone formation. This strategy improved vascularized bone regeneration and supports the potential of this biomaterial platform for repair of complex bone defects. A GO-SF/nHA scaffold co-delivers IL-4 and VEGF through biphasic sustained release, synergistically inducing macrophage M2 polarization and EPC angiogenesis to promote vascularized bone regeneration.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Drug Delivery and Translational Research
·
2026
·
IF 6.9
Rao Qing,Pan Binbin,Peng Zheng,Yu Qimin,Yu Yulong,Wang Huijun
Dysregulated immune responses and excessive inflammation are central drivers of early sepsis progression. Interleukin-10 (IL-10) is a key anti-inflammatory cytokine. However, the therapeutic potential and mechanistic basis of IL-10 mRNA delivery in correcting early immune dysfunction during sepsis remain poorly defined. In this study, we developed a mannose-modified mesoporous silica nanoparticle system (MMP/IL-10 mRNA) to stabilize IL-10 mRNA and enable sustained release for the immunoregulatory treatment of sepsis. The nanoparticles exhibited uniform size, structural stability, efficient mRNA protection, and sustained IL-10 expression. In LPS-stimulated macrophages, MMP/IL-10 mRNA enhanced cell viability, promoted M2 polarization, suppressed pro-inflammatory cytokine production (TNF-α, IL-1β, IL-6, and IL-17A), and restored the IL-10/IL-17A balance. In a cecal ligation and puncture–induced murine sepsis model, MMP/IL-10 mRNA improved 7-day survival, enhanced bacterial clearance, alleviated lung and liver injury, and ameliorated systemic inflammatory imbalance. Mechanistically, treatment reduced TLR4 expression, prevented IκBα degradation, inhibited NF-κB p65 phosphorylation, and upregulated endogenous IL-10 and DEL-1. Importantly, no detectable toxicity or organ damage was observed. These findings demonstrate that MMP/IL-10 mRNA enables efficient, sustained, and safe delivery of IL-10 mRNA, attenuates early hyper-inflammation, and improves immune dysregulation in sepsis, highlighting its potential as a promising immunoregulatory therapeutic strategy. Graphical abstract
The alternative text for this image may have been generated using AI.
✓ 已验证
DOI
PubMed
高影响因子
Biomaterials Research
·
2025
·
IF 9.6
Zheng Wang,Ying Chu,Yu Hu,Xue Fang,Jingyi Du,Mingshuang Li,Yuxin Zha,Jiabing Ran,Aixi Yu
Tendinopathy markedly impairs patients’ quality of life, yet effective therapies remain limited. Conventional therapeutic strategies have largely overlooked the pivotal role of macrophages (M φ ) and M φ -mediated immunoregulation in this condition. To address this, we designed a microneedle patch (MP) for the in situ delivery of rosmarinic acid (RosA), aiming to modulate local M φ phenotype. We evaluated the therapeutic efficacy of this RosA-MP against tendinopathy and investigated the underlying mechanism. Briefly, the RosA-MP was fabricated by incorporating RosA into a microneedle array composed of poly(hydroxyethyl methacrylate-co-3-acrylamidophenyl boronic acid-co-(3-methacrylamidopropyl)trimethylammonium chloride) [poly(HEMA-co-3APBA-co-MAPTAC)], which was covalently linked to a flexible filter paper matrix. Formation of phenylboronic acid ester bonds between the 3APBA moieties and RosA endowed the RosA-MP with enhanced stiffness and skin penetration capability, facilitating sustained RosA release. Furthermore, the MAPTAC moieties provided instant hydrophilic swelling upon skin insertion, accelerating RosA delivery via increased internal osmotic pressure. In vitro and in vivo assays demonstrated that the RosA-MP simultaneously induced in situ M 2 M φ polarization and scavenged reactive oxygen species. Mechanistic investigation revealed that suppression of the NLRP3 inflammasome contributed to the rationale behind M φ polarization. Using a collagenase type I-induced rat tendinopathy model, we assessed the therapeutic effects of the RosA-MP. Results confirmed that the RosA-MP effectively treated tendinopathy and exhibited significant advantages over injection therapy.
研究领域:
生物医学工程
·
免疫治疗
·
药物递送
·
再生医学
·
炎症研究
✓ 已验证
产品关联
DOI
PubMed
高影响因子
INTERNATIONAL IMMUNOPHARMACOLOGY
·
2026
·
IF 5.6
An Chang,Yi Wang,Zan Cui,Yuan-Yuan Liu,Ling-Qi Meng,Jia-Shi,Zhi-Yong Qin,Jia-Ning Gong
Pulmonary fibrosis (PF) is a progressive and lethal interstitial lung disease, characterized by excessive extracellular matrix deposition and architectural distortion of the lung parenchyma. Its pathogenesis involves interconnected pathological events, including dysregulated epithelial-mesenchymal transition (EMT), chronic inflammation driven by M2-polarized macrophages, and abnormal fibroblast activation. These intertwined mechanisms contribute to the limited efficacy of current anti-fibrotic therapies, which often fail to achieve lesion-specific targeting and disease reversal. Moreover, the clinical utility of existing drugs is further hampered by poor bioavailability and insufficient accumulation at fibrotic sites. To overcome these challenges, we developed a multifunctional nanotherapeutic platform, termed HPA@NPs, through co-assembly of hyaluronic acid-platycodin D (HA-PD) and aspirin-platycodin D (ASA-PD) conjugates. This nanosystem enables concurrent modulation of multiple PF-relevant pathological features. Physicochemical characterization showed that HPA@NPs possess uniform nanoscale size, low critical aggregation concentration, and excellent colloidal stability, supporting prolonged blood circulation. In vitro, HPA@NPs showed efficient cellular uptake consistent with their HA-based design and were accompanied by inhibition of EMT, fibroblast activation, and M2 macrophage polarization, together with reduced expression of IL-10 and Arg-1. In bleomycin-induced PF mice, HPA@NPs significantly improved pulmonary function and attenuated histopathological damage. At the molecular level, HPA@NPs down-regulated fibrosis-related markers (Col1a1, TGF-β1, α-SMA), while up-regulating the epithelial tight-junction protein ZO-1 and down-regulating mesenchymal markers (N-cadherin, vimentin), confirming effective reversal of EMT. The therapeutic outcome of HPA@NPs surpassed that of monotherapy, highlighting a synergistic anti-fibrotic effect. Importantly, HPA@NPs exhibited no detectable cytotoxicity, hemolytic activity, or major organ toxicity at therapeutic doses, demonstrating favorable biocompatibility. In summary, HPA@NPs integrate precise targeting, multi-pathway synergy, and excellent biosafety, offering a promising translational strategy for PF treatment.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CELLULAR AND MOLECULAR LIFE SCIENCES
·
2026
·
IF 6.5
Qu Qiao,Li Zhilong,Wang Dalu,Wu Di,Yin Hongzhuan
Colorectal cancer (CRC) remains a major contributor to cancer-related mortality worldwide. Although minichromosome maintenance protein 10 (MCM10) is known for its role in DNA replication, its involvement in CRC progression has not been fully elucidated. To identify key oncogenic drivers, RNA sequencing was performed on seven paired CRC and adjacent normal tissues, revealing significant upregulation of MCM10 (Log2FC = 2.359, P = 0.003). Public GEO datasets (GSE240623, GSE200427) further confirmed MCM10 overexpression in CRC tissues. This finding was validated in 40 paired clinical samples using qPCR and Western blotting. Prognostic relevance was assessed using hazard ratio (HR). Functional assays in vitro and in vivo were conducted to assess the effects of MCM10 on proliferation, migration, invasion, and M2 macrophage polarization. The m6A modification of MCM10 mRNA was evaluated through METTL3 and YTHDF1 interaction analysis in HCT116 cells. MCM10 was significantly upregulated in CRC tissues and associated with poor prognosis (HR = 1.54, P = 0.00077). MCM10 overexpression promoted CRC cell proliferation and invasion and was correlated with elevated M2 macrophage infiltration in tumor xenografts (6.16 ± 0.85% vs. 11.7 ± 1.13%, P < 0.05). Conditioned media from MCM10-overexpressing cells enhanced THP-1 macrophage polarization by increasing secretion of CCL2, CCL5, and IL10. Mechanistically, METTL3-mediated m6A modification stabilized MCM10 mRNA in a YTHDF1-dependent manner. MCM10 is aberrantly overexpressed in CRC and linked to worse clinical outcomes. Its oncogenic effects involve METTL3/YTHDF1-mediated mRNA stabilization and enhanced M2 macrophage polarization, suggesting a potential role in immune microenvironment modulation.
✓ 已验证
产品关联
DOI
高影响因子
Bioactive Materials
·
2023
·
IF 18.9
Peng Cheng,Xudong Xie,Liangcong Hu,Wu Zhou,Bobin Mi,Yuan Xiong,Hang Xue,Kunyu Zhang,Yuxiao Zhang,Yiqiang Hu,Lang Chen,Kangkang Zha,Bin Lv,Ze Lin,Chuanlu Lin,Guandong Dai,Yixin Hu,Tengbo Yu,Hankun Hu,Guohui Liu,Yingze Zhang
It is imperative to develop and implement newer, more effective strategies to address refractory diabetic wounds. As of now, there is currently no optimal solution for these wounds. Hypoxic human umbilical vein endothelial cells (HUVECs)-derived exosomes have been postulated to promote diabetic wound healing, however, its effect and molecular mechanism need further study. In this study, we aimed to investigate whether hypoxic exosomes enhance wound healing in diabetics. Based on our high-throughput sequencing, differentially expressed lncRNAs (including 64 upregulated lncRNAs and 94 downregulated lncRNAs) were found in hypoxic exosomes compared to normoxic exosomes. Interestingly, lncHAR1B was one of the prominently upregulated lncRNAs in hypoxic exosomes, showing a notable correlation with diabetic wound healing. More specifically, hypoxic exosomes were transmitted to surrounding cells, which resulted in a significant increase in lncHAR1B level, thereby relieving the dysfunction of endothelial cells and promoting the switch from M1 to M2 macrophages under high glucose conditions. Mechanistically, lncHAR1B directly interacted with the transcription factor basic helix-loop-helix family member e23 (BHLHE23), which subsequently led to its binding to the KLF transcription factor 4 (KLF4) and promoted KLF4 expression. In our in vivo experiments, the use of hypoxic exosomes-loaded HGM-QCS hydrogels (Gel-H-Exos) resulted in rapid wound healing compared to that of normoxic exosomes-loaded HGM-QCS hydrogels (Gel-N-Exos) and diabetic groups. Consequently, our study provides potentially novel therapeutic approaches aimed at accelerating wound healing and developing a practical exosomes delivery platform.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ACS Applied Polymer Materials
·
2023
·
IF 5
Yiwen Pan,Shaoqing Chen,Yanyan Meng,Mu He,Chun Liu,Cheli Wang,Xinye Ni
The treatment of bone defects caused by diseases, trauma, or tumor has always been a great clinical challenge. Implantation of bone biomaterials into bone defect areas is an effective method for bone injury repair. In this study, we used three-dimensional (3D) printing technology to prepare nano-hydroxyapatite (nHA)/sodium alginate (SA)/gelatin (Gel) hydrogel scaffolds loaded with different ratios (0, 0.13, 0.26, 0.39, 0.53, and 0.79‰) of emodin (EM) (EM/nHA/SA/Gel). Scanning electron microscopy showed that the scaffolds had a smooth surface without fracture and nHA was evenly distributed on the surface. The cell proliferation and migration results showed that the 0.39‰ EM group, in particular, could significantly promote the proliferation and migration of mouse embryonic osteoblast precursor (MC3T3-E1) cells and significantly increase the mRNA expression of osteogenic differentiation-related genes (bone morphogenetic protein/BMP-2, BMP-9, osteocalcin). In addition, the 0.39‰ EM group exhibited the best effect on osteogenic differentiation-related proteins (alkaline phosphatase, Runx 2, OSX). The expression of M2 polarization-related genes (arginase-1, CD206) also significantly increased after the treatment with the 0.39‰ EM group. Micro-CT showed that in the rat skull defect model, the EM/nHA/SA/Gel scaffold group significantly promoted bone regeneration after being implanted into the skull for 30 days. Our results indicate that the EM/nHA/SA/Gel hydrogel scaffolds can not only directly promote the proliferation and differentiation of osteoblasts but also indirectly promote osteogenic differentiation by supporting M2 polarization of macrophages. EM/nHA/SA/Gel hydrogel scaffolds are potential bone tissue engineering materials for bone regeneration.
✓ 已验证
产品关联
DOI
高影响因子
INTERNATIONAL IMMUNOPHARMACOLOGY
·
2023
·
IF 5.6
Yao Guo,Jing Cui,Xueyi Liang,Taoyu Chen,Chong Lu,Tao Peng
Pancreatic cancer (PC) is a serious threat to human health, with most patients diagnosed at the advanced stages of the disease. Treatment with gemcitabine (GEM) leads to PC GEM resistance. In addition, cancer stem cell (CSC)-derived exosomes play an important role in cancer progression. We aimed to investigate the role and mechanism of action of PC stem cell-derived exosomes in PC drug resistance and progression. CSC-derived exosomes increased the proportion of F4/80+/CD86 + cells and levels of M2 polarization factors. miR-210 is expressed in CSC-derived exosomes. Thus, following co-culture, miR-210 was taken up by macrophages. Transfection or the addition of miR-210 mimics increased the proportion of F4/80+/CD206 + cells and levels of M2 polarization factors. Further, the miR-210 targets inhibited the levels of FGFRL1. The FGFRL1 overexpression plasmid also inhibited miR-210-mediated M2 polarization. After co-culture of THP-M2 cells with PC cells and treatment with GEM, the survival rate, migration rate, and levels of MDR, YB-1, BCRP, p-PI3K, p-AKT, and p-mTOR in PC cells increased. And THP-M2 increased the tumor volume and MDR, YB-1, BCRP, p-PI3K, p-AKT, and p-mTOR levels. Overall, miR-210 from PC stem cell-derived exosome targets and inhibits FGFRL1 to promote macrophage M2 polarization, which activates the p-PI3K/p-AKT/p-mTOR pathway and increases GEM resistance.
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）